Glycidamide- 13C3

Glycidamide-13C3 Internal Standards in the Determination of Cascade Metabolites of Acrylamide

Zhang Y, et al. Journal of agricultural and food chemistry, 2020, 68(24), 6748-6758.

An ultra-high performance liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous analysis of cascade metabolites of acrylamide in rat and human urine, including acrylamide, glycidamide, N -acetyl-S -(2-carbamoylethyl)-l -cysteine (AAMA), and more.
Pretreatment process of urine samples
· Before analyzing the urine samples, a small portion (10 μL) of thawed rat urine was mixed with 10 μL of a solution containing various internal standards (acrylamide-d3, glycidamide-13C3, AAMA-d3, GAMA-d3 and iso-GAMA-d3, each at a concentration of 10 μg/mL).
· Following this, the samples were diluted with 980 μL of the original mobile phase solution (consisting of 2% acetonitrile in 0.1% formic acid) and vortexed for 30 seconds. The resulting solution was then filtered through a 0.22-μm millipore syringe filter and prepared for injection into the UHPLC-MS/MS system.
· In a similar manner, 1 mL of human urine was thawed, spiked with the internal standards, and treated with formic acid (10 μL) and ammonium formate (2 mL, 50 mmol/L, pH 2.5). After vortex mixing for 30 seconds and centrifugation at 10000 r/min for 5 minutes to remove any impurities, a two-step solid-phase extraction (SPE) process was utilized for further cleaning of the supernatant due to the presence of more complex impurities in the human urine matrix.